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Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
Salivary markers of systemic diseases ppt
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Salivary markers of systemic diseases ppt

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  • 1. SALIVARY MARKERS OF SYSTEMIC DISEASES
    BY
    Y.PRAGZNA(10T21S0113)
    UNDER THE GUIDANCE OF
    Mrs. M. SANDHYA M.PHARM
    ASSOCIATE PROFESSOR
  • 2. CONTENTS:
    • Biomarkers
    • 3. Saliva physiology
    • 4. Advantages & disadvantages
    • 5. Saliva collection, processing & storage
    • 6. Diagnostic applications
    • 7. Salivary proteome & transcriptome
    • 8. New technologies of measurement
  • BIOMARKERS:
    Biomarkers are defined as cellular, biochemical and molecular characteristics by which normal/abnormal processes can be recognized and/or monitored.
  • 9. SALIVARY MARKERS
    Non invasive diagnosis of disease and monitoring of general health
  • 10. SALIVA PHYSIOLOGY:
    • Saliva is the mixed product of 3 major salivary glands.
    Parotid
    Sub mandibular
    Sub lingual
    • It is slightly acidic(pH 6-7).
    • 11. Several physiological & pathological conditions modify saliva production quantitatively.
  • Functions of saliva
    PROTECTIVE
    FOOD & SPEECH RELATED
  • SALIVA AS A DIAGNOSTIC FLUID
    ADVANTAGES
    • It is safe, non-invasive and simple.
    • 21. No needles and No cringing.
    • 22. Home testing.
    • 23. It has an average protein concentration of 1.5-2 mg/ml.
    • 24. Cost of shipping & Storage tend to be lower.
    Informative analytes are present in lower amounts in saliva than in serum.
    DISADVANTAGES
  • 25. Whole saliva collection
    Ductal secretions collection
    Parotid secretions collection
    Sub mandibular & sub lingual secretions collection
    Saliva collection
  • 26. PROCESSING & STORAGE:
    All procedures must be performed on ice:
    (1)Divide the saliva samples into multiple 330μL samples placed in cryotubes at -80ºc.
    Further processed for
    • Protein analysis: Add protease inhibitors like aprotinin(0.33µL), 3.3µL PMSF.
    • 27. RNA analysis: Add 1.65µL of SUPERase inhibitor.
    (2)Invert gently to mix.
    (3)Store all fractions at -80ºc.
  • 28. Salivary/Oral Fluid Biomarkers Possibilities of Use
    DNA Standard genotyping
    Genetic information of the hosting human body
    Bacterial infection (oral microbes in the mouth) Diagnosing carcinomas of head & neck
    Forensics
    mRNA Information on transcription rates
    RNA Viral/bacterial identification
    Proteins Genetic information
    Translational regulation
    Diagnosing periodontitis
    Detecting dental cavities
    Immunoglobulin Diagnosing viruses
    Metabolites Diagnosing periodontitis
  • 29. DIAGNOSTIC APPLICATIONS OF SALIVA:
    • Drugs, hormones:
    • 30. Intra cellular pathways
    Active & Passive diffusion
    • Extra cellular pathways
    Ultra filtration
    • Serum components:
    • 31. Capillary barrier
    • 32. Interstitial spaces
    • 33. Cervicular fluid
    • 34. Membrane of acinar
    & ductal cells
  • 35.
    • SJOGRENS SYNDROME:
    There are raised concentrations of NaCl, IgA, IgG, lactoferrin, inflammatory mediators such as PGE2, TXB2, Interleukin-6 etc.
    • CYSTIC FIBROSIS:
    Saliva of CF patients contain increased calcium & phosphate levels, urea & uric acid and abnormally elevated levels of PGE2.
    • DIABETES MELLITUS:
    It effects flow rate & composition of saliva. Salivary alterations includes higher levels of IgA, Salivary Peroxidase, glucose content, K, Salivary total protein & amylase.
    • ORAL MUCOSAL DISEASES:
    Nitric oxide levels were increased.
  • 36.
    • ONCOLOGY:
    P53 antibody can be detected in oral squamous cell carcinoma, increased levels of salivary kallikrein levels in oral malignant tumors, CA 15-3 in breast cancer. CVS:
    Determination of salivary amylase activity before 6hrs after cv surgery.
     KIDNEY DYSFUNCTION:
    Hypo salivation , changes in taste, ammonium smelling breath, oral mucosal pain.
    EPILEPSY:
    Gingival hypertrophy, IgA deficiency
  • 37. VIRAL DISEASES:Acute hepatitis A & B were diagnosed based on the
    presence of IgM antibodies in saliva.HIV:Salivary IgA levels.BACTERIAL INFECTIONS:By monitoring the presence of antibodies to microorganisms found in saliva & oral cavity
  • 38.
    • AGAIN & AGAIN ABOUT TOBACCO:
    • 39. Smoking causes decrease in salivary important antioxidant levels.
    • 40. Salivary nicotine levels are considered to be the most reliable marker
  • Drug monitoring
    ANTI-ASTHMATICS:Theophylline
    ANTI EPILEPTICS: Carbamazapine, Diazepam, Ethosuximide, Lamotrigine, Topiramate, Phenytoin, Primidone, Valproic acid
    ANTI ARRYTHMIC/ANTI HYPERTENSIVES: Digoxin, Metoprolol, and Procainamide.
    ANTI MICROBIALS:Clarithromycin, Gentamycin, Isoniazid, Ofloxacin, Sulfanilamide
    ANTIPYRETICS: Acetaminophen, Antipyrine, Paracetamol
    ANTI NEOPLASTICS:Cisplatin, Methotrexate, Texol, Topotecan
    ANTI VIRAL: Quinine
    LITHIUM PSYCHOTROPIC:Lithium, Amitriptyline
    MISCELLANEOUS: Nicotine, Ethanol, Caffeine, Tolbutamide, Quinine
    ILLICIT DRUGS: Amphetamine, Barbiturates, Benzodiazepines, Opiods, Cocaine, Phencyclidine
  • 41. HORMONE MONITORING:
    Steroids like cortisol, aldosterone, testosterone, insulin
    SIALOCHEMISTRY ANALYSIS:
    Early monitoring of an exposure to lead, cadmium, mercury because of higher salivary levels of these elements
  • 42. CLASSIFICATION OF SALIVARY BIO MARKERS:
  • Diagnostic molecular targets
    SALIVARY PROTEOME:
    Use of ‘Divide & conquer’ strategy
    • Initial fractionation of proteins with separation techniques
    • 46. Digestion with proteolytic enzyme, trypsin
    • 47. Analysis by 1-D, 2-D LC-MS
    • 48. Finally data is processed & submitted for database searching
  • SALIVARY PROTEOMIC ANALYSIS
  • 49. SALIVARY TRANSCRIPTOME:
    • It includes mRNA molecules that cells use to convey the instructions carried by DNA for subsequent protein production.
    • 50. At present, the main strategy for identification of salivary transcriptomic biomarkers is through microarray technology.
  • SALIVARY TRANSCRIPTOME:
    DNA MICROARRAY TECHNOLOGY
  • 51. NEW TECHNOLOGIES FOR MEASURING SALIVARY BIOMARKERS:
    • POC DEVICE
    In this POC device, saliva(100-300μL) is placed into the salivary collection module, and then delivered into the Nano-biochip.
    • MICROBEAD ARRAY
    Micro pits within a silicon wafer are populated with a variety of chemically sensitized bead micro reactors.
  • 52. MICRO BEAD ARRAY TECHNIQUE
    NANO-BIOCHIP
  • 53. CONCLUSION:
    The components of saliva act as a “Mirror of the body’s health”,
    widespread use and growing acceptability of saliva as a
    diagnostic tool is helping individuals, researchers, health care
    professionals and community health programs to better detect
    and monitor diseases and to improve the general health of the
    public
  • 54. REFERENCES:
    • Mandel ID. A contemporary view of salivary research. Crit Rev Oral Biol Med 1993; 4:599.
    • 55. Raymond G, Schipper a, Erika Silletti a, Monique H, Vingerhoeds. Saliva as research material: biochemical, physiochemical, and practical aspects. Archives of Oral Biology 2007 ;( 52):1114-35.
    • 56. Richard P, Jiri S, Jana V, Edgar F, Petr M, Jindrich P et al. Saliva as a diagnostic medium. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2009; 153(2):103-10.
    • 57. Eliaz Kaufman .The diagnostic applications of saliva. A review. Crit Rev Oral Biol Med 2002; 13(2):197-212.
    • 58. www.med.kufauniv.com/ar/journals2010/res%20(1).pdf
    • 59. www.jofs.in/files/Artical-11.pdf
    • 60. www.linkgroup.hu/docs/pcs2/218-08CurrGen.pdf
    • 61. www.wikipedia.com
     
  • 62. THANK YOU
  • 63. QUERIES???

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